Research progress on sensitive genes related to ferroptosis

WU Hao-lin, XU Yi-qian, FANG Xing-yue, WANG Ai-ping, MENG Qing-wen, LIU Qibing,FAN Hao-fei

1.Key Laboratory of Brain Science Research and Transformation in Tropical Environment of Hainan Province, Haikou 571199, China

2.Department of Pharmacology, School of Basic Medicine and Life Sciences, Hainan Medical University, Haikou 571199, China

Keywords:

ABSTRACT

1.Introduction

Ferroptosis phenomenon was first discovered by Sonam et al in 2003, and formally named by Dixon in 2012, mainly a new form of procedural cell death catalyzed by abnormal lipid oxide metabolism in cells[1].In 2021, Yao et al found that Ferroptosis played an important role in neurological diseases such as cerebral hemorrhage,ischemic stroke, neurodegenerative diseases, and cardiovascular and cerebrovascular diseases[2]Among neuron Ferroptosis is closely related to neurological disease and hemorrhagic stroke,cerebral cytolysis causes hemoglobin release, hemoglobin further decomposed into heme and its oxidation form of heme chloride,induce Ferroptosis[2].Meanwhile, in cancer treatment studies,cancer cells grow a higher iron demand than normal cells, therefore,triggering Ferroptosis sensitive related genes, targeted induction of Ferroptosis has a good advantage[3].It can be speculated that cancer death can induce differentiated death and avoid normal cells.Second, inhibiting different Ferroptosis sensitive genes can reduce neuronal damage and protect the nervous system.Therefore, how to regulate Ferroptosis sensitive genes, effectively or directional induce or inhibit Ferroptosis, play a differentiated damage or protective role is particularly important.This paper mainly summarizes the known genes related to Ferroptosis sensitivity and its induction mechanism to provide reliable clues for Ferroptosis in disease treatment or prevention research.

2.Regulates Ferroptosis-related genes inducing Ferroptosis

Ferroptosis is genetically regulated, And showed the various metabolic changes, Depending on the protein metabolic pathway,Ferroptosis-related genes are divided into four categories: iron metabolism: ACO1, CISD1/mitto NEET, FANCD2, FTH 1, HMOX1,HSBP1, IREB2, NCOA4, NFS1, PHKG2, STEAP3, TFRC; Lipid metabolism: ACSF2, ACSL4, CS, GPX4, LOX, LPCAT3, PEBP1;Antioxidative metabolism: MRP1, CARS, CBS, GCLC, HMOX1,KEAP1, NOX, NQO1, NRF2, SLC7A11; Energy metabolism: G6PD,GLS2, GOT1, microRNA 137, SLC1A5, Its function and regulation effects are shown in Table 1.Currently recognized and most reported regulatory Ferroptosis sensitive related genes include inhibition of glutathione peroxidase 4 (GPX4), FSP1, SLC7A11, P53, etc[4].

Tab1 List of ferroptosis-related genes

2.1 The GPX4 gene

G PX4 protein is an important lipid repair enzyme in the human body, which plays an important function in producing lipid messenger molecular pathways, mainly responsible for the degradation of lipid peroxide, removing toxic intermediates, and is regarded as a key regulator for Ferroptosis[4].GPX4 protein resists both iron and oxygen-dependent lipid peroxidation by converting a lipid peroxide (LOOH) into a non-toxic lipid[4].Glutathione (GSH)An important intracellular antioxidant synthetic from glutamate,cysteine and glycine, is a cofactor and synthetic substrate of GPX4 protein, essential for GPX4 protein to play a lipid repair function[33].By inhibiting System XC-causing cysteine consumption and lack of GSH synthetic substrate, thereby weakening the antioxidant enzyme GPX4 protein function, eventually resulting in oxygen homeostasis imbalance in somatic cells, inducing Ferroptosis[34].Recent studies have shown that cancer recurrence from resistant cancer cells is closely related to Ferroptosis, and Ferroptosis occurrence can remove cancer cells that escape treatment, thus providing an effective strategy to prevent cancer recurrence[35].Among them, GPX4 is considered a sensitive gene and important target for resistant cancer cells through Ferroptosis killing, with inducers Erastin, Imidazole ketone erastin (IKE), azazpyridine[36-43], See Table 2.

After the death of iron, the expression of its sensitive gene GPX4 is necessarily downregulated.In 2019, Zou Y et al found in renal transparent cell cancer studies that HIF-2 through hypoxia-induced activation of lipid drop-related protein (HILPDA) expression,inhibited GPX4 gene expression, resulting in reduced GPX4 protein synthesis, promoting polyunsaturated lipid aggregation,thus increasing transparent cell cancer sensitivity to Ferroptosis[44].In 2021 Thayyullathil F et al.found that GSH scavenger Erastin inhibited System XC-, on the plasma membrane by reduced cell ability to acquire cysteine, reduced cysteine synthesis, and in turn triggered membrane lipid ROS accumulation and Ferroptosis[45].Furthermore, Lee N et al.found that another GPX4 inhibitor, RSL3,could also induce cell lipid oxidation and Ferroptosis[46].Wei Z et al.found that RSL3 (0.1 M) could significantly induce the death of human lens epithelial cells and lens epithelial iron in mice, both in vitro and in vitro[34].More research has shown that the Ferroptosissensitive gene GPX4, may target the Ferroptosis, providing reference for the research and development of targeted anti-tumor or protective cell drugs.

2.2 The FSP1 gene

FSP1 was originally known as mitochondrial Apoptosis Induction Inductive Factor 2 (Apoptosis-inducing factor mitochondrial 2,AIFM2) and was named due to the sequence similarity to apoptosisrelated factor AIFM1[47].FSP1 is an Ferroptosis sensitive related gene where overexpression in cells can significantly protect cells from Ferroptosis inductor factors[47].This is the first discovery of an enzyme catalytic system to compensate for GPX4 deficiency[48]In 2019, both German and American groups found that FSP1 cardamylation was critical to inhibit Ferroptosis[47,48].FSP1 protein prevents lipid oxidation by reducing the coenzyme-Q10 (CoQ10),thereby inhibiting Ferroptosis[48].In 2020, Wu Warren found that FSP1 and GPX4 proteins were involved in non-interference inhibition, which used CRISPR-Cas9 screening to identify Ferroptosis suppression protein FSP1 as an effective Ferroptosis resistance factor[49].Data show that cartmeg acylation raises FSP1 into the mass film, becomes redox, reduces CoQ10 levels,generates lipophilic free radical capture antioxidants (RTA) and avoiding lipid peroxidation[48].The Researchers further found that FSP1 gene expression was positively associated with Ferroptosis resistance in hundreds of cancer cell lines and found resistance to Ferroptosis in FSP1 protein-mediated cultured lung cancer cells and tumor allografts in mice[47].Thus, these data determine that FSP1 is a critical component of the non-mitochondrial CoQ antioxidant system, independently promoting Ferroptosis occurrence based on the GSH classical GPX4 pathway.To explore other inhibitors of Ferroptosis, Zhang et al used the CRISPR/Cas9 method for genome-wide screening, finding that when the FSP1 gene was knocked out, the tumor cell resistance to RSL3 disappeared and the degree of Ferroptosis increased[50].It is shown that pharmacological targeting of FSP1 gene and GPX4 inhibitors induce Ferroptosis[48].In summary, FSP1/CoQ10/NAD (P) H acts as a single parallel induction system works in collaboration with GPX4 and GSH to inhibit phospholippid peroxidation (pLPO) and Ferroptosis.

2.3 The SLC7A11 gene

SystemXC-encoded by the SLC7A11 gene belongs to the family of isodimic amino acid transporters, has the function of exchanging L-cysteine and L-glutamate and is considered to be one of the major signaling pathways associated with Ferroptosis[51].Glusteine is exchanged at 1: 1, so glutamate levels affect systemXCfunction[52].Extracellular high concentrations of glutamate inhibit SystemXC-and induce Ferroptosis.Mice knocking off systemXCcan prevent neurotoxic damage due to reduced extracellular glutamate levels[53].Erastin can act as an iron-promoting death trigger, inhibiting SystemXC-function and consuming GSH, by eventually inactivating GPX4, causing lipid peroxide accumulation,which further leads to an increase in ROS[53].Cyclic iron trivalent iron ion (Fe3+) Introducing cells through the ferrin receptor (TFR),then converted into Fe2+in the inner body [54].Fe2+Overdose causes lipid ROS accumulation through Fenton reaction, resulting in turn Ferroptosis[54].

2.4 The P53 gene

Tumor suppressor protein p53 (TP53) is a stress response transcription factor capable of activating multiple target genes under different stress conditions, including oxidative stress, genotoxic damage, oncogene activation, and hypoxia[55].p53 gene expression can significantly stimulate biological functions such as cell cycle block, apoptosis, aging, metabolism, and autophagy regulation[28].p53 protein was found in the 1970 s and plays a key role in cells to various stress responses including DNA damage, hypoxia,nutritional hunger, and oncogene activation[56].In 2015, XX first found that p53 increased sensitivity to Ferroptosis by inhibiting SLC7A11 expression, and[57].Li S et al.showed increased P53 expression in 1-methyl-4-phenylpyridine (MPP) induced Ferroptosis cell models, resulting in reduced 7 members of the solute carrier family 11 (SLC7A11) and GPX4 expression[28].Hong T et al found Ferroptosis in mutant ovarian cancer, the main mechanism may be p53-dependent downof SLC7A11 expression[58].

Studies have shown that p53 protein activation inhibits SLC7A11 transcription, resulting in reduced cysteine intake, reduced intracellular GSH, increased ROS levels, and lipid peroxidation[28].Thus, increased ROS-induced Ferroptosis sensitivity in p53-activated cells[28].The p53 mutant (p53 KR) cannot induce cell cycle block or apoptosis, and can still can induce Ferroptosis, indicating that the effect of p53 on Ferroptosis is multiple[57].Moreover, Erasatininduced Ferroptosis is not prevented by inhibiting autophagy,apoptosis, or necrosis, suggesting that p53 regulates cysteine uptake to promote Ferroptosis as an effective mechanism[58].

2.5 Others

At present, the study of Ferroptosis related genes such as ACO1,ACSF2, MRP 1 and G6PD is less reported, and the main mechanism is likely to be related to Erastin.Although it may be involved in the occurrence of Ferroptosis, the mechanism of Ferroptosis is unknown and remains to be further verified.

3.Outlook

Research is important in disease treatment.A growing number of basic research has found that inducing or blocking Ferroptosis can affect the occurrence and development of multiple diseases,providing a potential target for treatment, especially for diseases resistant to conventional drugs[59].Take drug resistant cancer,inducing cancer cell death is the therapeutic purpose of many chemotherapy and biotargeted drugs, and cancer cells are prone to form resistance or resistance that drugs cannot kill.But it was found that the resistant cancer cell GPX4 gene is sensitive to Ferroptosis and is targeted to induce cancer cell death[60].Therefore, targeting Ferroptosis provides a new perspective for the treatment of various diseases.

4.Summary

This review mainly addresses the impact of the Ferroptosissensitive related genes of GPX4, FSP1, SLC7A11 and P53 on the development of Ferroptosis.There is a partial understanding of the morphological, biological, and mechanical pathways of cell Ferroptosis, and the different Ferroptosis-sensitive related genes of different cell types.But the trigger process of Ferroptosis sensitive genes involves various mechanisms, the precise regulation of signaling pathway, Ferroptosis and disease, whether and other cell death jointly mediated the progress of the disease, therefore,according to pathological characteristics, targeted trigger Ferroptosis sensitive related gene experimental model, targeted in-depth study of Ferroptosis mechanism, study its role in different disease types,looking for the relevant disease treatment target and targeted drug research and development is of important significance.

Tab2 Summary of GPX4 Inactivation or Loss

Note: VDAC: membrane voltage-dependent anion channel; NF-κB: nuclear factor B, TGF-: transformation growth factor- β,COX-2: epoxoxidase-2; Raf-1:pro-oncoges serine / threonine-protein kinase, B-Raf: serine / threonine rat-filtered sarcoma carcinogenic homologous B1 Gene, FIN56: Ferroptosis Inducer.

Inducers Target mark Function References Mazolone Mostetin An effective, metabolic stability, selective SystemXC-inhibitor, and Ferroptosis inducer [36]AlazSulpyridine An effective, specific NF-B TGF-and COX-2 inhibitor,Inducing Ferroptosis [37]Elastin Induced Ferroptosis by acting on the mitochondrial VDAC [38]Solafini Multiple kinase inhibitors of Raf-1 and B-Raf that activate Ferroptosis [39]Butsteine Sulfonimide Glutamine-cysteineAcid Ligase Lower the cell GSH content [40]Artesunate Glutathione S-Transferase Inhibitors of STAT-3 and output protein 1 (EXP1) that induce Ferroptosis [41]BAY 87-2243 Dehydrogenase Suppresses mitochondrial complex I activity, triggering elevated dependent mitochondrial ROS, resulting in Ferroptosis [42]FIN56 Shalene Synthase Binding and activating Shalene Synthase to inhibit non-steroid metabolites-most likely Coenzyme Q10, -enhanced cell sensitivity to FIN56-induced Ferroptosis [43]

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